Objective To analyze the reasons of misdiagnosis of primary syphilitic chancre and strengthen the understanding of atypical features of this disease.Methods A case series of twenty-seven challenging primary syphilis patients who were not immediately recognized as chancre was included in our study.The clinical data including the patients\' age,sex,skin lesions,HIV status,syphilis serologic test results,treatment,and follow-up results were collected.Hematoxylin-eosin and immunohistochemistry staining of skin biopsy sections were reviewed.Results Four female cases with extragenital chancres presenting as erythema or erosive skin lesions on the nipple were misdiagnosed as Paget\'s disease or eczema.The disorder of missed or misdiagnosed male cases manifested as syphilitic balanitis or multiple chancres on the penis root and adjacent pubis rather than coronal sulcus or frenum.Patients with nonreactive nontreponemal tests at initial presentation were also easily missed or misdiagnosed.Conclusion Primary syphilis presenting as multiple lesions rather than a single chancre,at atypical locations,or with a nonreactive nontreponemal test result,tends to be missed or misdiagnosed.